Purification and biochemical characterization of phytase from Bacillus cereus isolated from gastrointestinal tract of African giant snail (Achatina fulica) / Purificación y caracterización bioquímica de fitasa de Bacillus cereus aislada del tracto gastrointestinal de caracol gigante africano (Achatina fulica)

Phytases are specialized enzymes meant for phytic acid degradation. They possess ability to prevent phytic acid indigestion, including its attendant environmental pollution. This study was aimed at investigating biochemical properties of purified phytase of B. cereus isolated from Achatina fulica. Phytase produced from Bacillus cereus that exhibited optimal phytate degrading-ability of all the bacteria isolated was purified in a three-step purification. The biochemical properties of the purified enzyme were also determined. The phytase homogeny of approximately 45 kDa exhibited 12.8-purification fold and 1.6% yield with optima phytate degrading efficiency and maximum stability at pH 7 and 50 °C. Remaining activity of 52 and 47% obtained between 60 and 70 °C after 2 h further established thermostability of the purified phytase. Mg2+ and Zn2+ enhanced phytate hydrolysis by the enzyme, while Na+ showed mild inhibition but Hg2+ severely inhibited the enzymatic activity. Km and Vmax were estimated to be 0.11 mM and 55.6 μmol/min/mL, displaying enzyme-high substrate affinity and catalytic efficiency, respectively. Phytase purified from Bacillus cereus, isolated from African giant snails, has shown excellent characteristics suitable for phytic acid hydrolysis and could be employed in industrial and biotechnological applications.(AU)

Purification and biochemical characterization of phytase from Bacillus cereus isolated from gastrointestinal tract of African giant snail (Achatina fulica).

Phytases are specialized enzymes meant for phytic acid degradation. They possess ability to prevent phytic acid indigestion, including its attendant environmental pollution. This study was aimed at investigating biochemical properties of purified phytase of B. cereus isolated from Achatina fulica. Phytase produced from Bacillus cereus that exhibited optimal phytate degrading-ability of all the bacteria isolated was purified in a three-step purification. The biochemical properties of the purified enzyme were also determined. The phytase homogeny of approximately 45 kDa exhibited 12.8-purification fold and 1.6% yield with optima phytate degrading efficiency and maximum stability at pH 7 and 50 °C. Remaining activity of 52 and 47% obtained between 60 and 70 °C after 2 h further established thermostability of the purified phytase. Mg2+ and Zn2+ enhanced phytate hydrolysis by the enzyme, while Na+ showed mild inhibition but Hg2+ severely inhibited the enzymatic activity. Km and Vmax were estimated to be 0.11 mM and 55.6 µmol/min/mL, displaying enzyme-high substrate affinity and catalytic efficiency, respectively. Phytase purified from Bacillus cereus, isolated from African giant snails, has shown excellent characteristics suitable for phytic acid hydrolysis and could be employed in industrial and biotechnological applications.

Biochemical studies of enzyme-induced browning of African bush mango (Irvingia gabonensis) fruit pulp.

The purpose of this study was to examine the biochemical properties of African bush mango (Irvingia gabonensis) pulp PPO. PPO was purified from I. gabonensis fruit pulp in three steps and characterized. A purification fold of 343 with specific activity of 216 U/mg and 13% recovery were obtained as well as molecular weight of 32.67 kDa was observed. The optimum pH and temperature were found to be pH 7.0 and 50 °C respectively while the enzyme showed instability at low pH 2-4 with total inactivation at pH 2 but maximal at pH 5-9 with remaining residual activity of 60-90%, whereas, total enzyme activity inactivation was observed at 90 °C. However, Cu2+, Fe2+ and Mg2+ enhanced the PPO activity but inhibited by Ca2+, Ba2+, K+ and Na+. Notably, purified PPO was inactivated completely by urea at concentration above 10 mM while Km and Vmax values were estimated to be 7.34 mM and 0.36 U/min for catechol, 10.76 mM and 0.30 U/min for L-DOPA, and 14.90 mM and 0.26 U/min for tyrosine, respectively. The activity of PPO in I. gabonensis fruit and its juicy product could be controlled at high temperature in acidified medium.

Xeronine structure and function: computational comparative mastery of its mystery.

Morinda citrifolia (Noni) fruit has a long history of dietary use in tropical regions of the world. Pharmacological properties that have been attributed to the fruit include anti-inflammatory, anti-cancer, and antioxidant properties. Xeronine, a small alkaloid which has been patented (US4543212) is one of the bioactive compounds of Noni fruit, which is believed to be capable of modifying the molecular structure of specific inactive proteins thereby regulating proper folding to active enzymes. Despite reports of the potential of Xeronine as therapeutic agent, its presence is controversial and its structure has not been explored. In this study, standard chemoinformatics tools and servers such as ChemSketch, ChemMine, Swisstargetprediction, SwissADME and Swisssimilarity have been employed to predict its possible structure. In addition, synthetic xeronine structures based on the known bioactive components of Noni fruit were designed. Results showed that the hypothetical structure of xeronine provided by the patent inventor is a mystery based on its <5% probable protein targets and no similarity match to the US Food and Drug Administration (FDA) approved drugs and experimental compounds by in silico evaluation. By constrast, final designed xeronine structure possess all the features that were described in the patent document, and has >40% probable protein targets related to neurodegenerative diseases such as Alzheimer's disease (AD), which possibly justifies the key function stated in the patent.

Optimization and prediction of antioxidant properties of a tea-ginger extract.

A response surface approach was used to investigate the effects of temperature, concentration, and time on the antioxidant properties (total flavonoid (TF), total phenol (TP), peroxide scavenging activity (PS), iron chelating activity (IC), DPPH radical-scavenging ability (DPPH), ABTS assay (ABTS)) of aqueous extract of tea-ginger (2:1) powder. Color indices, pH, and redox potential of the tea-ginger powder were also measured and used as independent variables for the prediction of antioxidant properties of the extract using ordinary least square (OLSR), principal component (PCR), and partial least square (PLSR) regression. The R (2) values for TP, TF, ABTS, and PS response surface models were 0.8873, 0.9639, 0.6485, and 0.5721, respectively. The OLSR, PCR, and PLSR were able to provide predictive models for DPPH, TP, and TF of the tea-ginger extract (P < 0.05). The PLSR gave the most parsimonious model with an R (2) of 0.851, 0.736, and 0.905 for DPPH, TP, and TF, respectively.